Revealing phylogeographic structure is important for accurate subspecies delineation and understanding a species’ evolutionary history. In leopards (Panthera pardus), there are currently nine subspecies recognized. On the African continent, only one subspecies occurs (P. p. pardus), although mitochondrial DNA from historical samples suggests the presence of three putative continental clades: (1) West Africa (WA), (2) Central Africa (CA), and (3) Southern Africa (SA). So far, whole genome data did not recover this phylogeographic structure, although leopards in the southern periphery of their distribution range in Africa have not yet been investigated in detail. The Mpumalanga province of South Africa is of particular interest, as here, the CA and the SA clade possibly meet. The aim of this study was to characterize the first mitogenomes of African leopards from Mpumalanga, to help clarifying how South African leopards fit into continental patterns of genetic differentiation. Complete mitogenomes from nine leopards, including a strawberry leopard, were assembled de novo and included in phylogenetic analysis, in combination with 32 publicly available mitogenomes. Bayesian inference and maximum likelihood analyses identified two deeply diverged putative clades within South Africa, which were more genetically distinct than two subspecies in Asia. The clades dated back to 0.76-0.86 million years ago, indicating that they originated during the climatically unstable Mid-Pleistocene, as seen in other large mammals. The Pleistocene refuge theory states that the maintenance of savanna refugia in East and Southern Africa promoted the divergence between populations. As such, leopards may reflect the unique climatic history of southern Africa, which has resulted in eminent and endemic genetic diversity.